Kumar, Swati Ashok (2015) Developing a physiologically relevant blood brain barrier model for the study of drug disposition in glioma. Doctoral thesis, University of Central Lancashire.
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The blood brain barrier is a highly selective physical, transport and metabolic barrier, that limits the disposition of drugs to the CNS; a pertinent problem to the successful treatment of glioma. There has been an emergence of cell based in vitro models as potential tools to predict the in vivo permeation of therapeutics at the BBB, however most of the available models have only been characterised for just the physical barrier and not the metabolic barrier comprised of efflux transporters and drug metabolising enzymes. In addition, a review of all the available models found that there was a lack of multicellular models cultured using all human components. There was an urgent need to develop a model which is physiologically relevant and focusses on the metabolic barrier present at the blood and brain junctions with a goal to delivering chemotherapeutics to CNS tumours. The aim of the study was to design an all human, fully characterised and physiologically relevant in vitro BBB model for the prediction of permeability of drugs across the blood brain barrier to glioma. Cell cultures investigated included the glioma cell lines (U87MG, 1321N1), non-cancerous cell lines (SVGp12, hCMEC/D3), glioma short term cultures (BTNW914, BTNW370) human endothelial culture (HBMEC), human astrocytes (HA) and human pericytes (HBVP). The cell cultures were characterised and studied for the expression of activity levels for efflux transporters; ABCB1 and ABCG2 and drug metabolising enzymes; CYP3A4 and CYP2D6. The culturing conditions of cells was optimised by using human serum, human fibronectin to maximise the expression and activity of these proteins. The cell showing high expression and activity were used to design the mono-, co- and tri- culture in vitro BBB model on a transwell insert. The models were assessed on the basis of TEER, tight junction protein expression and expression and activity of efflux transporter and DMEs. These models were also constructed on 3D Alvetex scaffolds and exposed to shear stress by dynamic perfusion. Finally model was used to screen permeability of novel lipid NPs and a glioma specific targeting aptamer SA43.The human primary cultures (HBMEC, HBVP and HA) were selected to model the BBB and the tri-culture model gave the highest TEER of 258 Ω/cm2. The tri-culture model on alvetex scaffold with perfusion showed a TEER of 769 Ω/cm2. The screening of NPs found the docetaxel and curcumin lipid NPs were permeable through the barrier. In future, the developed BBB model will be further characterised and used commercially for screening of novel glioma therapeutics and drug delivery systems.
|Item Type:||Thesis (Doctoral)|
|Uncontrolled Keywords (separate with ;):||BBB modelling; Glioma therapeutics; Invitro BBB models|
|Schools:||Faculty of Clinical & Biomedical Sciences > School of Pharmacy and Biomedical Sciences|
|Deposited By:||Paul Harrison|
|Deposited On:||29 Jul 2016 14:46|
|Last Modified:||22 Feb 2017 09:50|
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